Standardization of Bindu Ghrita

[Find the meaning and references behind the names: Bindu, Pal, Vega, Choudhary, Kalpana, Sneha, Meena, Geeta]

Research Article This work is licensed under a CC BY 4.0 License International Research Journal of Ayurveda & Yoga Vol. 5 (8),61-69, August, 2022 ISSN: 2581-785 X https://irjay.com/ DOI: 10.47223/IRJAY.2022.5808 Standardization of Bindu Ghrita Geeta Choudhary 1 ,Mohar Pal Meena 2 , Sakhitha K.S. 3 ,Trupti Naikare 4 1 MD scholar , Dept. of Rasashastra and Bhaishajya Kalpana National Institute of Ayurveda , Jaipur (Rajasthan) 2 Associate Professor, Dept. of Rasashastra and Bhaishajya Kalpana National Institute of Ayurveda, Jaipur (Rajasthan) 3 Assistant Professor, Dept. of Rasashastra and Bhaishajya Kalpana National Institute of Ayurveda , Jaipur (Rajasthan) 4 MD scholar , Dept. of Rasashastra and Bhaishajya Kalpana, National Institute of Ayurveda , Jaipur (Rajasthan) INTRODUCTION The fundamental objective of Ayurveda is the maintenance of health i.e. “ Swasthasya Swasthaya Rakshanam ” and curing diseases i.e “ Aturasya Vikara Prashamanam ” 1 . Bhaishajya Kalpana is one of the branch of Ayurveda that deals with the pharmaceutical preparation of Ayurveda formulations. Sneha Kalpana is one of the unique dosage forms of Ayurveda which is explained in Bhaishajya Kalpana . Sneha Kalpana is medicaments prepared from Sneha i.e. lipid material. Sneha siddhi is the process of making Article Info Article history: Received on:28-06-2022 Accepted on: 19-08-2022 Available online: 31-08-2022 Corresponding author- Geeta Choudhary, National Institute of Ayurveda, Jaipur Rajasthan – 302002 Email;geetchoudhary 800@gmail.com ABSTRACT: One of the major contributions of Ayurveda is the dosage form called Sneha Kalpana where ghrita and t aila are processed in such a way that all the phytochemicals from the plant parts are transferred to the lipid media, mostly the ones which are lipid soluble. The process of making Sneha Kalpana is divided into two steps; the first is Sneha Murchchhana and the second is Sneha Paka . Sneha Murchchhana is performed to remove doshas from the lipid media while Sneha Paka is the preparation of medicated ghrita or taila ( Sneha Kalpana ). The product obtained after the process of Sneha paka is known as Siddha ghrita or Siddha taila . There are various formulations of Siddha ghrita and Siddha taila given in the Ayurveda text. Bindu Ghrita is one of the formulations of ghrita , mentioned in Ayurveda texts like Chakradatta , Sharangadhara Samhita , Bhaishajya Ratnavali , Gadanigraha and Yoga Ratnakara . The word Bindu suggests its therapeutic use indicating the number of Bindus applied on the Nabhi ( umbilicus ) which is equal to the number of Virechana Vega ( purgations ). Internal use of Bindu Ghrita is also suggested in some texts. Bindu Ghrita is used to treat Udara rogas mainly in Jalodara (Ascites). Its other indications are as Kushtha , Aadhmana , Mandaagni , Gulma etc. There have been very few works done on this formulation, and no work on the standardization of Bindu Ghrita has been undertaken till date. This study focuses on the pharmaceutical as well as analytical standardization of Bindu Ghrita as per the reference of Sharangadhara Samhita and is one of the first studies reported on its standardization. Keywords – Sneha Kalpana , Bindu Ghrita , Jalodara , Virechana Vega , Murchchhana , Udara roga .

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Geeta et. al “Standardization of Bindu Ghrita .” : 2022; 5 (8):61-69 62 oleaginous medicaments. While preparing Sneha Kalpana we have to consider three factors, these are Kalka dravya , Sneha dravya and Drava dravya . Kalka dravya is the paste of herbal ingredients, Sneha dravya is any lipid (Usually taila and ghrita ) and Drava dravya could be Ksheera , Kwatha or water i.e. liquid media. These three are taken in a definite proportion and are subjected to a heating pattern which is unique for the Sneha siddhi process. Lipid soluble drugs have better Pharmacokinetic action in Sneha Kalpana form compared to Kwatha or other watersoluble dosage forms because lipid-soluble substances readily permeate into the cell membrane due to their fatsoluble nature. Bindu Ghrita is one of the Sneha Kalpana , which is mentioned in texts like Chakradatta , Sharangadhara Samhita , Bhaishajya Ratnavali , Gadanigraha and Yoga Ratnakara . This ghrita is named Bindu because of its therapeutic dose which causes its unique action. Local application of one bindu of Bindu Ghrita on Nabhi will result in one Virechana Vega and so on, i.e. a number of bindu will cause an equal number of Virechana Vegas . This unique property is responsible for the name Bindu . One Bindu is approximately about 0.5 ml 2 Bindu Ghrita is indicated in a broad spectrum of diseases, mostly the ones caused by the malfunctioning of Agni . Diseases like Udara roga , Kushtha , Adhmana , Mandagni and Gulma etc. are therapeutic indications of Bindu Ghrita . Till date no studies about the Standardization of Bindu Ghrita were carried out, therefore considering the wide array of therapeutic activities of Bindu Ghrita and the unavailability of data on standardization of the same, it was found necessary to standardize Bindu Ghrita . Considering all the above-mentioned factors the present study was planned accordingly. MATERIALS & METHODS The study was mainly carried out in two different stages • Pharmaceutical Study • Analytical Study [A] Pharmaceutical Study The pharmaceutical study consists of 1. Preparation of coarse powder of ingredients 2. Ghrita Murchchhana 3. Preparation of Bindu Ghrita (3 batches) 1. Preparation of coarse powder of ingredients Collection of Raw Materials - All the raw materials were procured from the pharmacy of the National Institute of Ayurveda, Jaipur. Except for Shankhini ( replaced with Shankhapushpi ), Satala ( Var. Euphorbia tirucalli ), Dantiphala ( Jayapala ), Koshataki , Devdali , Nilini , Aparajita , Hemkshiri were procured from raw drugs supplier from Maharashtra and fresh Matulunga , Snuhi Ksheera , Arka Ksheera were procured from a local gardener from Jaipur and identified by the expert of Dravyaguna department of NIA Jaipur. Before undergoing any pharmaceutical process there was the need for careful inspection of raw materials to detect foreign matters like sand, dust and seeds etc. The foreign materials were separated by the hand picking and sieving process. Then they were stored in an airtight glass container and used as per requirement. Physical impurities were removed from the herbal drugs and drugs were sun-dried and coarse powder was made using laboratory-based Mortar and Pestle. The Bindu Ghrita was prepared using the reference mentioned in Sharangadhara Samhita . Three samples of Bindu Ghrita were prepared in the Drug Manufacturing Laboratory of the Department of Rasashastra and Bhaishajya Kalpana , NIA Jaipur. Physico-chemical study and Analytical study were performed in the Drug Testing Laboratory of NIA Jaipur and S.R. laboratory Jaipur. (Ayush certified laboratory) 2. Ghrita Murchchhana The reference used for Ghrita Murchchhana was taken from Bhaishajya Ratnavali , Principle followed in the process was Sneha Paka principle itself. Table 1 Showing Ingredients of Ghrita Murchchhana 3 Out of all the drugs mentioned in table 1, coarse powder of drugs numbered 2-7 were taken and added with Matulunga Nimbu Swarasa (Fresh Fruit Juice) and mixed properly in a mortar pestle. A required quantity of water was added till the Kalka was properly formed. After the formation of Kalka , Go-ghrita (2.5 Kg) was heated to make it free from moisture. The temperature achieved during the process was around 100˚C, this heating process took 20 mins to make the entire amount of ghrita free from moisture. After this go-ghrita was kept aside for self-cooling. When the temperature of go-ghrita was around 70˚C, Kalka was added to it. Kalka was added slowly in the ghrita and stirred continuously. After this process mentioned quantity of water (10 litres) was added in this. The heating was started and was continued till the Sneha Siddhi Lakshanas were achieved. Heating was discontinued once ghrita started showing all Sneha Siddhi Lakshanas 4 Ghrita was kept aside for self-cooling (70˚C) and was filtered through a double-layered Muslin cloth. For

[Find the meaning and references behind the names: Shanti, Long, Brown, Ama]

Geeta et. al “Standardization of Bindu Ghrita .” : 2022; 5 (8):61-69 63 all three batches of Bindu Ghrita , the required amount of Murchchhita go-ghrita was prepared in one batch. The final product i.e. Murchchhita ghrita ( 2328 g with 6.88% loss ) was weighed and kept in a PET jar once it was at room temperature. (Fig. 1) 3. Preparation of Bindu Ghrita The reference used in the preparation of Bindu Ghrita is from Sharangadhara Samhita Madhyam Khanda , Sneha Kalpana Adhyaya . The principle used in this process was Sneha Paka . Table 2 Showing Ingredients of Bindu Ghrita for each sample 5 Drugs numbered from 2 - 20 in table 2 were cleaned and yavakut churna i.e. coarse powder was prepared. The process was done separately for each ingredient. For making Kalka , coarse powder of all drugs was added with a small quantity of water to achieve a Kalka (Paste) like appearance. Later Murchchita go-ghrita was taken into a stainless - steel vessel and it was heated just to make it moisture free. (Temperature achieved 100˚C) Then Kalka was added slowly with continuous stirring. After the addition of Kalka , ghrita temperature was noted around 70˚C. Later to this water, Snuhi Ksheera and Arka Ksheera were added in the mentioned quantity. The temperature was again fallen down to 50˚C. The mixture was heated till all the water was evaporated. This was indicated by the appearance of Sneha Siddhi Lakshanas . Phena Shanti , the appearance of Varti and absence of cracking sound in the Ghrita and Kalka when exposed to flame are the Sneha Siddhi Lakshana . After the appearance of these Lakshana , the heat was discontinued. After a few minutes, the prepared Bindu Ghrita was filtered using a double-layered Muslin cloth and stored inside a PET jar after self-cooling. (Fig. 2) In a similar way, a second sample and the third sample of Bindu Ghrita were prepared. The three samples were coded as BG-1, BG-2 and BG-3 respectively in this study. Table 3 Showing the weight loss of 3 Samples of Bindu Ghrita in Sneha Paka [B] Analytical Study Tests were carried out as per the protocol for testing ASU drugs which included Organoleptic parameters, Physicochemical parameters like Viscosity, Specific gravity etc. and other tests like Heavy metal analysis, Aflatoxins & Microbial analysis. HPTLC of the samples was also carried out. Organoleptic Test – Table 4 Results of Organoleptic characters of BG-1, BG- 2 and BG-3 Physico-chemical and Miscellaneous Test 6 – Table 5 Results of the Physico-chemical and Miscellaneous parameters of Bindu Ghrita Samples RESULTS AND DISCUSSIONS Ayurveda formulations like Bindu Ghrita are not yet standardized in API and AFI. There was an immense need to standardize this formulation using Physico-chemical or Analytical parameters The word Bindu suggests its therapeutic use indicating the number of bindus (drops) applied on the Nabhi ( umbilicus ) which is equal to the number of Virechana Vega (purgations). Bindu Ghrita one among the rare formulations were external application of the medicament can cause purgations in the patients probably due to a combination of powerful purgative drugs like Danti, Jayapala, Hemakshiri, Snuhi Ksheera etc. Internal use of Bindu Ghrita is also suggested in some texts. Bindu Ghrita is used to treat Udararogas mainly Jalodara (Ascites). Its other indications are Kushtha , Aadhmana , Mandaagni , Gulma etc. In the present study, Bindu Ghrita was prepared according to the reference of Sharangadhara Samhita. Murchchhana was done prior to Sneha paka to remove Ama dosha which can be considered as unwanted components in the raw ghrita , like intermediate chemical constituents, dissolved gases, adulterants and moisture present in raw ghrita or developed due to long time storage. During Sneha paka Mandagni (i.e. 60-80˚C) was maintained throughout the procedure in order to prevent the evaporation of active principles, to avoid adhering of Kalka on the bottom of the vessel as well as to provide sufficient time for the active principles to get incorporated into ghrita . Phena Shanti , absence of frothing is one of the Sneha Sidhi Lakshanas which denotes completion of the process of Sneha paka. Ghrita is a saturated fatty acid with a single bonded structure. On heating, it undergoes hydrolysis. Once the water content evaporates it results in subsiding of froth in the ghrita . Absence of crackling sound and appearance of Varti indicate the absence of moisture content in the prepared ghrita. In the present study an average loss of 9.33% was observed which may be probably due to the absorption of ghrita by the kalka part. Normally the colour of ghrita depends on the constituents present in the ghrita , the colour of all three samples was found yellowish brown. The odour was typical aromatic which was seen in all three samples. After paka , the odour of samples leads to aromatic due to the addition of Kalka

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Geeta et. al “Standardization of Bindu Ghrita .” : 2022; 5 (8):61-69 64 dravyas and Drava dravyas . Minimal variations were found in the Physico-chemical parameters of all the three samples. Viscosity was in the range 35.75 cP to 37.50 cP, Specific gravity was found to be 0.9450 to 0.9595, Refractive index was 1.480 to 1.485, Iodine value between 27.80 to 30.75, Acid value found in the range 0.04 to 0.06, Peroxide value was between 3.50 to 3.98, Saponification value came out between 273.96 to 296.50, Ester value was 273.92 to 296.45 and the total fatty matter was 96.75 to 97.75. Rancidity was absent in all three samples. Heavy metals & aflatoxins found in all three samples were within permissible limits. Microbial analysis shows that all three samples have microbial count within normal limits. HPTLC report of samples BG-1, BG-2 & BG-3 show a large number of spots, which denotes that samples have a large number of Phytoconstituents. (Fig.3) This indicates the presence of Phytoconstituents in all multiple ingredients added in the preparation of Bindu Ghrita . HPTLC provides qualitative information on the main constituents of the drug. In the present study, methanolic extract of the sample was used and the solvent system was toluene and ethyl acetate in the ratio of 9:1 respectively. For determination of the TLC plate methanolic H 2 SO 4 was used. Samples of Bindu Ghrita were analyzed on different wavelengths 254 nm, 366 nm and 510 nm. BG-2 shows the maximum number of spots in visible light. The presence of these spots could be due to the addition of various Kalka dravyas and Drava dravyas . But due to the absence of any standards, the specific compound could not be separately identified.The scanning data shows different spots visualized get confirms the chemical nature and distribution pattern in the specified mobile phase.Table 6 Results of HPTLC Finger Print data of BG-1, BG-2 and BG-3 CONCLUSION The Physico-chemical parameters are important for the quality control of ASU formulations. The results of this study may be used as reference standards for Bindu Ghrita and will also be helpful in further studies of similar kind. Acknowledgements - National Institute of Ayurveda Conflict of interest - None Source of finance & support – National Institute of Ayurveda. ORCID Geeta Choudhary , https://orcid.org/ 0000-0001-9193-9192 REFERENCES 1 Shastri K,Charaka, Charaka Samhita, Viddyotini Hindi Commentory Sutrasthana 30, Shloka 26, Chaukhambha Viddya Bhavan Varanasi,2009.pp.587 2 Ayurvedic formulary of India, Part-1, 2 nd ed, National institute of scienve communication and information resources, CSIR. New delhi. pp.383 3 Shastri A.D Bhaishajya Ratnavali, Chapter Jwara Chikitsa- 5/1266 Choukambha Sanskrit samsthan, varanasi, 18 th edition, 2005. 4 Shastri A.D Bhaishajya Ratnavali, Choukambha Sanskrit samsthan, varanasi, 18 th edition, 2005 5 Murthy K,R Sharangadhara, Sharangadhara samhita, Madhyam khanda, 1 st edition Chapter 9, Shloka 13-14. Choukambha orientalia, Varanasi, 1984. 6 Anonyms, The Ayurvedic Pharmacopeia of India, Part-I, Vol.–VI, Department of Aayush, Ministry of Health and Family Welfare, Govt. of India, edition 2008. How to cite this article: Choudhary G, Meena M.P, Sakhitha K.S Naikare T “Standardization Of Bindu Ghrita ” IRJAY.[online]2022;5(8); 61-69 Available from: https://irjay.com DOI linkhttps://doi.org/10.47223/IRJAY.2022.5808

[Find the meaning and references behind the names: Raja, Seed, Krishna, Shweta]

Geeta et. al “Standardization of Bindu Ghrita .” : 2022; 5 (8):61-69 65 Table 1 Showing Ingredients of Ghrita Murchchhana S.No Name Latin Name Part Use Quantity 1 Go-ghrita Clarified Butter - 2.5 Kg 2 Haritaki Terminelia chebula Fruit 156.25 g 3 Vibhitaki Terminelia bellerica Fruit 156.25 g 4 Amalaki Embelica officinalis Fruit 156.25 g 5 Musta Cyprus rotandus Rhizome 156.25 g 6 Haridra Curcuma longa Rhizome 156.25 g 7 Matulanga Nimbu Citrus medica Swarasa (Fresh Fruit Juice) 156.25 g 8 Water - - 10 Ltr. Table 2 Showing Ingredients of Bindu Ghrita for each sample S.No. Name of Dravya Latin Name Part Use Quantity 1 Go-ghrita ( Murchchhita ) Clarified Butter - 768 g 2 Chitraka Plumbago zeylanica Root 12 g 3 Shankhapushpi Convolvulus pluricaulis Panchanga 12 g 4 Haritaki Terminalia chebula Fruit 12 g 5 Kampillaka Mallotus philipinesis Phala raja 12 g 6 Shweta Trivrita Operculina terpethum Root 12 g 7 Krishna Trivrita Operculina terpethum Root 12 g 8 Vriddhadaru Argyreia nervosa Root 12 g 9 Aragvadha Casia fistula Fruit Pulp 12 g 10 Danti Baliospermum montanum Root 12 g 11 Dantiphala Croton tiglium Seed 12 g 12 Koshataki Luffa acutangular Fruit 12 g 13 Devdali Luffa echinate Fruit 12 g

Geeta et. al “Standardization of Bindu Ghrita .” : 2022; 5 (8):61-69 66 14 Nilini Indigofera tinctoria Whole Plant 12 g 15 Aparajita Clitoria ternatea Whole Plant 12 g 16 Satala Euphorbia tirucalli Whole Plant 12 g 17 Pippalimoola Piper longum Root 12 g 18 Vidanga Embelia ribes Fruit 12 g 19 Kutaki Pichrorrhiza kurroa Root 12 g 20 Hemkshiri Argemona maxicana Whole Plant 12 g 21 Snuhi Ksheera Euphorbia nerifolia Latex 288 g 22 Arka Ksheera Calotropis procera Latex 96 g Table 3 Showing the weight loss of 3 Samples of Bindu Ghrita in Sneha Paka Sample Date of Commencement Date of Completion Initial weight of Murchchhita Ghrita Weight of Bindu Ghrita after Paka Loss of Ghrita % Loss Mean of Loss ( in % ) BG-1 27/01/2022 29/01/2022 768 g 683.52 g 84.48 g 11.00% 9.33 % BG-2 05/02/2022 07/02/2022 768 g 708.48 g 59.52 g 7.75% BG-3 11/02/2022 13/02/2022 768 g 696.58 g 71.42 g 9.30% Table 4 Results of Organoleptic characters of BG-1, BG-2 and BG-3 S.N Organoleptic Character Normal Go - Ghrita Murchchhita Goghrita SAMPLE-1 (BG-1) SAMPLE-2 (BG-2) SAMPLE-3 (BG-3) 1 Colour Light yellow Yellowish ( Dark Yellow ) Yellowish Brown Yellowish Brown Yellowish Brown 2 Odour Characteristic of Go-ghrita Characteristics of Kalka dravya specially Haridra Aromatic Aromatic Aromatic 3 Consistency Semisolid Semiliquid Liquid Liquid Liquid 4 Appearance Oily Oily Oily Viscous Oily Viscous Oily Viscous

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Geeta et. al “Standardization of Bindu Ghrita .” : 2022; 5 (8):61-69 67 Table 5 Results of the Physico-chemical and Miscellaneous parameters of Bindu Ghrita Samples S. NO Parameters BG – 1 BG – 2 BG – 3 1 Viscosity 35.75 cP 37.50 cP 36.75 cP 2 Specific gravity 0.9595 0.9450 0.9545 3 Refractive Index 1.480 1.485 1.482 4 Rancidity Absent Absent Absent 5 Iodine Value 28.50 27.80 30.75 6 Acid Value (mg KOH/g) 0.06 0.04 0.05 7 Peroxide Value 3.50 3.98 3.95 8 Saponification Value (mg KOH/g) 274.52 273.96 296.50 9 Ester Value 274.46 273.92 296.45 10 Total fatty matter 96.75 96.95 97.75 11 Heavy Metals (mg/Kg or ppm) Lead (Pb) Arsenic (As) Cadmium (Cd) Mercury (Hg) 1.08 0.21 BLQ (LOQ 0.01) BLQ (LOQ 0.01) 1.38 0.16 BLQ (LOQ 0.01) BLQ (LOQ 0.01) 0.93 0.17 BLQ (LOQ 0.01) BLQ (LOQ 0.01) 12 Total Aflatoxins ( mg/Kg or ppm ) Aflatoxin B 1 Aflatoxin B 2 Aflatoxin G 1 Aflatoxin G 2 BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) BLQ ( LOQ 0.05 ) 13 Microbial Analysis Total Bacterial Count Total Fungal Count <10 cfu/gm <10 cfu/gm <10 cfu/gm <10 cfu/gm <10 cfu/gm <10 cfu/gm Table 6 Results of HPTLC Finger Print data of BG-1, BG-2 and BG-3 Sample 254 nm 366 nm 510 nm BG – 1 16 spots visualize 9 spots visualize 10 spots visualize BG – 2 14 spots visualize 8 spots visualize 13 spots visualize BG – 3 14 spots visualize 10 spots visualize 10 spots visualize

Geeta et. al “Standardization of Bindu Ghrita .” : 2022; 5 (8):61-69 68 Fig. 1 Ingredients of Ghrita Murchchhana Pharmaceutical Process of Ghrita Murchchhana Fig. 2 Ingredients of Bindu Ghrita

Geeta et. al “Standardization of Bindu Ghrita .” : 2022; 5 (8):61-69 69 Pharmaceutical Process of Bindu Ghrita Fig. 3

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